Key Laboratory of Biology and Genetic Improvement of Horticultural Crops, Ministry of Agriculture, Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing 100081, China.
Institute of Horticulture, Henan Academy of Agricultural Sciences, Zhengzhou 450002, China.
Int J Mol Sci. 2022 May 5;23(9):5147. doi: 10.3390/ijms23095147.
Microspore culture, a type of haploid breeding, is extensively used in the cultivation of cruciferous crops such as cabbage. Heat shock (HS) treatment is essential to improve the embryo rate during the culture process; however, its molecular role in boosting early microspore embryogenesis (ME) remains unknown. Here we combined DNA methylation levels, miRNAs, and transcriptome profiles in isolated microspores of cabbage '01-88' under HS (32 °C for 24 h) and normal temperature (25 °C for 24 h) to investigate the regulatory roles of DNA methylation and miRNA in early ME. Global methylation levels were significantly different in the two pre-treatments, and 508 differentially methylated regions (DMRs) were identified; 59.92% of DMRs were correlated with transcripts, and 39.43% of miRNA locus were associated with methylation levels. Significantly, the association analysis revealed that 31 differentially expressed genes (DEGs) were targeted by methylation and miRNA and were mainly involved in the reactive oxygen species (ROS) response and abscisic acid (ABA) signaling, indicating that HS induced DNA methylation, and miRNA might affect ME by influencing ROS and ABA. This study revealed that DNA methylation and miRNA interfered with ME by modulating key genes and pathways, which could broaden our understanding of the molecular regulation of ME induced by HS pre-treatment.
小孢子培养是一种单倍体繁殖技术，广泛应用于十字花科作物如白菜的培育。热休克（HS）处理对于提高培养过程中的胚胎率至关重要；然而，其在促进早期小孢子胚胎发生（ME）方面的分子作用尚不清楚。在这里，我们结合了白菜 '01-88' 的分离小孢子在 HS（32°C 24 小时）和常温（25°C 24 小时）下的 DNA 甲基化水平、miRNA 和转录组谱，以研究 DNA 甲基化和 miRNA 在早期 ME 中的调控作用。两种预处理的全基因组甲基化水平有显著差异，鉴定出 508 个差异甲基化区域（DMR）；DMR 的 59.92%与转录物相关，miRNA 基因座的 39.43%与甲基化水平相关。值得注意的是，关联分析表明，31 个差异表达基因（DEGs）被甲基化和 miRNA 靶向，主要涉及活性氧（ROS）响应和脱落酸（ABA）信号转导，表明 HS 诱导了 DNA 甲基化，miRNA 可能通过影响 ROS 和 ABA 来影响 ME。本研究表明，DNA 甲基化和 miRNA 通过调节关键基因和途径干扰 ME，这可以拓宽我们对 HS 预处理诱导 ME 的分子调控的理解。
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